Journal: bioRxiv

Article Title: Porous membrane electrical cell-substrate impedance spectroscopy for versatile assessment of biological barriers in vitro

doi: 10.1101/2023.06.26.546641

Figure Lengend Snippet: (A) Gold leaf sheets were bonded to polyethylene terephthalate porous membranes using a pressured thermal bonding process, followed by photolithographic patterning and etching to form electrodes. (B) Fabrication of a custom 6-well “Transwell-like” insert incorporating PM-ECIS. (C) Application of electrical insulating masks on both sides of the porous membrane electrodes. (D) Fabrication of a custom 96-well device incorporating PM-ECIS. (E) Photograph of 6-well plate with three 6-well PM-ECIS inserts with flat flex cable gold finger connectors.

Article Snippet: For cell culture, patterned PM-ECIS electrodes were cut using a laser cutter (VLS 3.5, Universal Laser Systems) for integration into custom-made 6-well inserts and 96-well devices.

Techniques:

Journal: bioRxiv

Article Title: Porous membrane electrical cell-substrate impedance spectroscopy for versatile assessment of biological barriers in vitro

doi: 10.1101/2023.06.26.546641

Figure Lengend Snippet: (A) Representative traces of 4 kHz PM-ECIS measurement of human umbilical vein perivascular cells (HUVECs) initially seeded at densitites of 250,000 cells/cm 2 (red lines) or 100,000 cells/cm 2 (blue lines) in 96-well PM-ECIS devices. Dotted lines show traces from a separate device under the same conditions. Black lines are cell-free control devices. (B) PM-ECIS resistances normalized to cell-free controls across the three measured frequencies (400, 4000, 40000 Hz) at end of day 4 for the high (red) and low (blue) density cultures. (C) Representative traces of 4 kHz PM-ECIS resistances measured for HUVECs cultured in 6-well devices with either a single electrode (blue) or 6-finger electrode (red). Dotted lines show traces from a separate device under the same conditions. Black lines are cell-free control devices. (D) PM-ECIS resistances normalized to cell-free controls across the three measured frequencies (400, 4000, 40000 Hz) at end of day 4 for the 6-well single electrode devices with (blue) and without (black) HUVECs. N = 3 devices.

Article Snippet: For cell culture, patterned PM-ECIS electrodes were cut using a laser cutter (VLS 3.5, Universal Laser Systems) for integration into custom-made 6-well inserts and 96-well devices.

Techniques: Cell Culture

Journal: bioRxiv

Article Title: Porous membrane electrical cell-substrate impedance spectroscopy for versatile assessment of biological barriers in vitro

doi: 10.1101/2023.06.26.546641

Figure Lengend Snippet: Human umbilical cord perivascular cells (HUVECs) grown in 6-well devices were treated with serum-containing medium hour 10 (grey arrows) and monitored continuously by PM-ECIS. The network model developed by Giaver and Keese (1991) was applied to derive (A) cell-cell pericellular resistance, R b ; (B) cell-electrode pericellular resistance, α; and C) cell layer membrane capacitance, C m . (D) 4kHz PM-ECIS resistance measurements of HUVECs treated a t=0 (grey arrow) with media only (HUVECs) or media + thrombin (HUVECs + thrombin) to cause barrier distruption. (E) Calculated pericellular resistance, R b , for thrombin-disrupted HUVEC layer.

Article Snippet: For cell culture, patterned PM-ECIS electrodes were cut using a laser cutter (VLS 3.5, Universal Laser Systems) for integration into custom-made 6-well inserts and 96-well devices.

Techniques: